the whole blood from fish cultured in C. pyrenoidos green water for 15 days
Total RNA of whole blood specimens was extracted using TRIzolTM reagent (Invitrogen, USA) according to the manufacturers protocol. The RNA quality was assessed by Agilent 2100 Bioanalyzer (Agilent Technologies, USA) and RNase free agarose gel electrophoresis. The eukaryotic mRNA was enriched by Oligo(dT) beads and fragmented into short fragments using fragmentation buffer. The short RNA fragments were then reverse transcribed to synthesize cDNA libraries with random primers. The second-strand of cDNA was synthesized and purified with QiaQuick PCR extraction kit (Qiagen, The Netherlands). The cDNA was then ligated to Illumina sequencing adapters and the ligation products were sequenced on Illumina NovaSeq 6000 system (Gene Denovo Biotechnology Co., China).